Evaluation of the humoral immune response in patients receiving cryopreserved human amniotic membrane transplantation on the ocular surface
Lauriana Solecki Dr1,2,3, Jean-Baptiste Baudey Dr4,5, Christophe Picard Dr4,5, Bastien Mathéaud Dr1, Pauline Jamain Dr6, Isabelle Jollet Dr6, Lucas Hubert Dr4,5, Alain Coaquette Dr7, Adeline Desanlis Dr8, Xavier Lafarge Dr9,10, Pascal Pedini Dr4,5, Florelle Gindraux2,11.
1Service d’Ophtalmologie, Centre Hospitalier Universitaire de Besançon, F-25000 Besançon, France; 2UR 4662 Soins Intégrés, Nanomédecine, IA et Ingénierie pour la Santé (SINERGIES), Université Marie & Louis Pasteur, F-25000, Besançon, France; 3Service d’Ophtalmologie, Nouvel Hôpital Civil CHRU Strasbourg, F-67000 Strasbourg, France; 4Laboratoire d’immunogénétique, Établissement Français du Sang Provence Alpes Côte d’Azur-Corse, F-13005 Marseille, France; 5CNRS, EFS, ADES UMR 7268, Aix-Marseille Université, F-13005 Marseille, France; 6Laboratoire d’Histocompatibilité, Établissement Français du Sang Nouvelle-Aquitaine, F-86012 Poitier, France; 7Service de virologie, Centre Hospitalier Universitaire de Besançon, F-25000 Besançon, France; 8Activité d’ingénierie cellulaire et tissulaire, Établissement Français du Sang Bourgogne Franche-Comté, F-25000 Besançon, France; 9Laboratoire d’ingénierie tissulaire et cellulaire, Établissement Français du Sang Nouvelle-Aquitaine, F-33075 Bordeaux, France; 10INSERM U1035 Biothérapie des Maladies Génétiques Inflammatoires et Cancers, Université de Bordeaux, F-33075 Bordeaux, France; 11Service de Chirurgie Maxillo-Faciale, Stomatologie et Odontologie Hospitalière, Centre Hospitalier Universitaire de Besançon, F-25000 Besançon, France
Introduction: Human amniotic membrane (hAM) is considered to have low immunogenicity since Akle et al. (Lancet) reported no anti-HLA antibodies (Abs) in four volunteers after subcutaneous grafting of fresh hAM in 1981. But detection methods’ sensibility has since improved significantly (Luminex Single Antigen). Furthermore, hAM is a massive local input of allogenic HLA-G which potential immunogenicty has never been assessed. We aim to evaluate hAM immunogenicity in a clinical setting after ocular surface transplantation using modern sensitive techniques.
Objective: To assess the occurence of anti-HLA class I, II, and anti HLA-G Abs at 1 and 3 months after hAM transplantation.
Material and methods: We conducted a clinical trial at the University Hospital of Besançon (France) in collaboration with three French Blood Transfusion Centers (Ethical approval: 2023-A01447-38). Twenty-three patients were included between August 2022 and September 2023. Two patients had combined conjunctival and corneal indications. Of 23 patients, 9 had a history of immunizing events prior to hAM transplantation (pregnancies or transfusions).
Sera were collected on transplantation date and at 1 and 3 months post-transplant (M0, M1, M3).
Anti-HLA class I, and II Abs were assessed by flow cytometry using the gold standard Luminex Single Antigen technique. A mean fluorescence intensity (MFI) > 500 was considered positive. Anti HLA-G antibodies were assessed using an ELISA-based in house detection assay, derived from the MAIPA assay.
Results and discussion: Despite using a highly sensitive detection technique, neither de novo immunization nor change in the profile of classical anti-HLA Abs were observed at M1 or M3 compared to pre-transplant profiles. No anti-HLA-G Abs were identifed before or after hAM transplant.
Conclusion: Given the advances in the knowledge of HLA expression by the hAM and in detection sensitivity, confirming previous results in a larger cohort was important. This is the first study to investigate anti-HLA class I, II, and anti HLA-G Abs in a clinical setting after cryopreserved hAM transplantation using sensitive techniques. Our results corroborate and update those obtained in the 1980s. We demonstrate the absence of immunization post hAM transplantation, confirming its immunological safety and supporting the excellent clinical tolerance observed over several decades across various indications.
Appels d’offres Recherche 2023, Agence de la Biomédecine, France - Call for proposals 2023, French Agency of Biomedicine .
[1] Tolerance
[2] Allograft
[3] Human amniotic membrane
[4] Immunization
[5] Immunogenicity
[6] HLA-I, HLA-II, HLA-G
[7] Luminex, MAIPA